pCMV human IRS-1 CAAX
(Plasmid #11238)

• Depositing Lab: Ronald Kahn
• Publication: Kriauciunas et al Mol Cell Biol. 2000 Sep . 20(18):6849-59.

Backbone

• Vector backbone: pCMV histidinol
• Backbone size w/o insert (bp): 7583
• Vector type: Mammalian Expression
• Selectable markers: histidinol

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): XL1 Blue
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: IRS-1
• Alt name: Insulin Receptor Substrate 1
• Alt name: IRS1
• Species: H. sapiens (human)
• Insert Size (bp): 5800
• Mutation: C-terminal prenylation motif makes IRS-1 localize to the cell membrane.
• GenBank ID: NM_005544
• Entrez Gene: IRS1 (a.k.a. HIRS-1)
• Tag / Fusion Protein:
  • CAAX (C terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: SacI (not destroyed)
• 3′ cloning site: SalI (not destroyed)
• 5′ sequencing primer: CMV-F

Resource Information

• Articles Citing this Plasmid:
  • 2 References

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
• Industry Terms:
  • Not Available to Industry

Depositor Comments

pCMV human IRS-1 was cut with ApaLI and SalI. A 5' IRS-1 oligo at bp4746 (amino acid 1242) and 3'IRS-1 oligo with 3' IRS-1 sequence fused to the p21ras CAAX prenylation motif (CMSCKCVLS) were used for PCR and the insert was ligated into pCMV human IRS-1. Select cells expressing plasmid with approximately 5mM histidinol (Sigma), depending on the cell type.